The intrinsic factor-vitamin B12 receptor and target of teratogenic antibodies is a megalin-binding peripheral membrane protein with homology to developmental proteins.
The present report shows the molecular characterization of the rat 460-kDa epithelial glycoprotein that functions as the receptor facilitating uptake of intrinsic factor-vitamin B12 complexes in the intestine and kidney. The same receptor represents also the yolk sac target for teratogenic antibodies causing fetal malformations in rats. Determination of its primary structure by cDNA cloning identified a novel type of peripheral membrane receptor characterized by a cluster of eight epidermal growth factor type domains followed by a cluster of 27 CUB domains. In accordance with the absence of a hydrophobic segment, the receptor could be released from renal cortex membranes by nonenzymatic and nonsolubilizing procedures. The primary structure has no similarity to known endocytic receptors but displays homology to epidermal growth factor and CUB domain proteins involved in fetal development, e.g. the bone morphogenic proteins. Electron microscopic immunogold double labeling of rat yolk sac and renal proximal tubules demonstrated subcellular colocalization with the endocytic receptor megalin, which is expressed in the same epithelia as the 460-kDa receptor. Furthermore, megalin affinity chromatography and surface plasmon resonance analysis revealed a calcium-dependent high affinity binding of the 460-kDa receptor to megalin, which thereby may mediate its vesicular trafficking. Due to the high number of CUB domains, accounting for 88% of the protein mass, we propose the name cubilin for the novel receptor.
Moestrup
S K
SK
Department of Medical Biochemistry, University of Aarhus, 8000 Aarhus C, Denmark. skm@biobase.dk
Kozyraki
R
R
Kristiansen
M
M
Kaysen
J H
JH
Rasmussen
H H
HH
Brault
D
D
Pontillon
F
F
Goda
F O
FO
Christensen
E I
EI
Hammond
T G
TG
Verroust
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D011992:Endosomes / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000235:genetics; D004847:Epithelial Cells / Q000737:chemistry; D026541:Heymann Nephritis Antigenic Complex; D007150:Immunohistochemistry; D007437:Intrinsic Factor / Q000378:metabolism; D007672:Kidney Cortex / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013723:Teratogens / Q000378:metabolism; D014805:Vitamin B 12 / Q000378:metabolism; D015017:Yolk Sac / Q000737:chemistry
D000906:Antibodies; D019485:Bone Morphogenetic Proteins; D018076:DNA, Complementary; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D013723:Teratogens; C029486:intrinsic factor-cobalamin receptor; D004815:Epidermal Growth Factor; D007437:Intrinsic Factor; D014805:Vitamin B 12